Trem2 H157Y homozygous mice (Trem2H157Y/H157Y) were crossed with 5xFAD mice (The Jackson Laboratory, stock # 34848) to obtain the 5xFAD; Trem2H157Y/+ offspring. 5xFAD; Trem2H157Y/+ mice were used to setup breeding cages to establish the littermate cohorts with three genotypes including 5xFAD; Trem2+/+, 5xFAD; Trem2H157Y/+, and 5xFAD; Trem2H157Y/ H157Y. The genotype of 5xFAD mice was characterized through probe-based qPCR with the protocol provided by the Jackson Laboratory. All the 5xFAD mice used as breeders or in our experimental cohorts were hemizygous.
5xfad mice
Manufactured by Jackson ImmunoResearch
Sourced in United States, Montenegro
5xFAD mice are a transgenic mouse model that overexpresses five familial Alzheimer's disease (AD) mutations. This model is used in research to study the pathogenesis of Alzheimer's disease.
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Lab products found in correlation
5xfad, by Jackson ImmunoResearch (4 mentions)
Wild type c57bl 6 mice, by Jackson ImmunoResearch (3 mentions)
C57bl 6j, by Jackson ImmunoResearch (3 mentions)
C57bl 6 mice, by Jackson ImmunoResearch (2 mentions)
Fvb nj, by Jackson ImmunoResearch (2 mentions)
Wt c57bl 6 mice, by Jackson ImmunoResearch (2 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (2 mentions)
App psen1, by Jackson ImmunoResearch (1 mentions)
Tg6799, by Jackson ImmunoResearch (1 mentions)
C57bl 6jolahsd, by Inotiv (1 mentions)
Icr mice, by Orient Bio (1 mentions)
Mmrrc stock 34840, by Jackson ImmunoResearch (1 mentions)
C57bl 6, by Inotiv (1 mentions)
Pregnant cd1 mice, by Charles River Laboratories (1 mentions)
Tamoxifen, by Merck Group (1 mentions)
C57bl 6 substrain characterization panel, by Jackson ImmunoResearch (1 mentions)
Dereg mice, by Jackson ImmunoResearch (1 mentions)
Nestlet, by Ancare (1 mentions)
C57bl 6j control, by Jackson ImmunoResearch (1 mentions)
3xtg ad mice, by Jackson ImmunoResearch (1 mentions)
70 protocols using 5xfad mice
1
Generating 5xFAD; Trem2 Mutant Mice
2
Evaluating siRNA Silencing of APOE in AD Mice
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All experimental studies involving animals were approved by the University of Massachusetts Medical School Institutional Animal Care and Use Committee (IACUC Protocols #A‐2411 and #A‐1744) and performed according to the guidelines and regulations therein described. APP/PSEN1 (MMRRC 34832‐JAX) mice were bred in house and obtained from Jackson Lab. 5xFAD mice (MMRRC Stock No: 34840‐JAX) were obtained from Jackson Lab (experiments were performed on hemizygote APP/PSEN1 and 5xFAD mice). Initial aged APP/PSEN1 mouse colonies were a gift from Dr. Arya Biragyn from the National Institute on Aging.
To determine ideal sample size, we performed a power calculation using published data in transgenic mice, and preliminary data evaluating efficacy of the siRNAs used in this study. In published data, the genetic removal of Apoe (100%) in transgenic AD models resulted in greater than 90% pathologic improvement, and ≈50% Apoe silencing with ASOs results in ≈10% reduction in pathology. As shown in Alterman et al.,
24 (link) the Apoe‐targeting siRNAs silence > 95% of APOE in the brain. Thus, we based the calculation off a predicted mean reduction in amyloid burden of 50% after treatment with siRNAs targeting Apoe, and a standard deviation of 25%. To achieve 95% power with an alpha level of 0.01, 9 animals would be required per group. The complete power calculation is included in TableS1 .
To determine ideal sample size, we performed a power calculation using published data in transgenic mice, and preliminary data evaluating efficacy of the siRNAs used in this study. In published data, the genetic removal of Apoe (100%) in transgenic AD models resulted in greater than 90% pathologic improvement, and ≈50% Apoe silencing with ASOs results in ≈10% reduction in pathology. As shown in Alterman et al.,
24 (link) the Apoe‐targeting siRNAs silence > 95% of APOE in the brain. Thus, we based the calculation off a predicted mean reduction in amyloid burden of 50% after treatment with siRNAs targeting Apoe, and a standard deviation of 25%. To achieve 95% power with an alpha level of 0.01, 9 animals would be required per group. The complete power calculation is included in Table
3
Fat-1 transgenic mice for omega-3 research
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The fat-1 transgenic mouse can endogenously produce omega-3, due to expression of the fat-1 gene that converts omega-6 into omega-3 [28 (link)]. The C57BL/6 mice used here as the control (genetic background) group of fat-1 mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA). Four-month-old male fat-1 (n = 8) and C57BL/6 (n = 10) mice were used. The 5XFAD mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA) and were cross-bred with male 5XFAD mice and female B6SJL/F1 mice to obtain littermates. The 5XFAD transgenic mice possess both APP mutations (V717I, I716V, K670N/M671L) and PSEN1 mutations (M146L, L286V), which result in characteristic Aβ overexpression. Mice had freely available food and water and were bred in a space with a 12/12 h dark/light cycle. The number of animals was calculated in accordance with resource equation [68 ,69 ,70 (link)]. Animals were raised according to the National Institutes of Health (NIH) guidelines for the care and use of laboratory animals (NIH Publications No. 8023, revised 1978), and all experiments were approved by the Institutional Animal Care and Use Committee at Konyang University (P-18-03-A-01, approved on 01 August 2018).
4
Generation and Validation of 5xFAD Mice
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Experimental procedures with mice were approved by the Consejo Superior de Investigaciones Científicas (CSIC) animal ethics committee and the Autonomous Government of Madrid, in compliance with the Spanish and European Union guidelines. Control C57BL/6JOlaHsd mice were purchased from ENVIGO (Barcelona, Spain). Double transgenic mice in C57BL/6 J genetic background expressing under the control of the Thy1 promoter both mutant human Aβ precursor protein 695 (APP695) with the Swedish (K670N, M671L), Florida (I716V), and London (V717I) FAD mutations and human presenilin 1 (PS1) harboring the M146L and L286V FAD mutations (Tg6799 or 5xFAD mice) [35 (link)] were purchased from The Jackson Laboratory (Bar Harbor, ME) (strain #008,730). 5xFAD mice were genotyped as indicated by The Jackson Laboratory. A h5xFAD mouse strain was obtained after repeated inbreeding of originally hemizygous mice. h5xFAD mice always yielded genetically homogeneous litters when crossed with WT mice (n = 20 litters). Homozygosity was confirmed by qPCR genotyping using specific primers of the human APP transgene (upstream primer: 5′-AGGATGGTGATGAGGTAGAG-3′ (from NM_201414 sequence) and downstream primer complementary to: 5′-CTGCTGTTGTAGGAACTCGA-3′ (896–915 bp and 1011–1030 bp from the NM_201414 sequence, respectively)), with the Kcn3a genomic sequence (see below) as a reference.
5
Transgenic 5XFAD Mouse Model of Alzheimer's
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Transgenic mouse (strain name; B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V) 6799Vas/Mmjax) carrying five mutations associated with early onset familial Alzheimer’s disease (FAD) was used in the experiment. The 5XFAD mice were obtained from Jackson Laboratory (USA) and have been maintained by mating with C57BL/6 X SJL wild type mice. Institute of Cancer Research (ICR) mice (strain name; Crl:CD1, male, six-week-old) were purchased from Orientbio Inc. (Seoul, Korea). The strain is a fertile albino mouse that is widely used for the disease modeling studies. All mice were bred in a laboratory animal breeding room at Yonsei University (Seoul, Korea). They were housed in groups of five per cage with a controlled temperature, humidity, and a 12/12 hour light/dark cycle. Water and food were available ad libitum. All animal experiments were carried out in accordance with the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals. The research protocol was approved by the Institutional Animal Care and Use Committee of Yonsei University, Seoul, Korea (IACUC-A-201806-744-01).
6
Accelerated Alzheimer's Mouse Model
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C57BL/6 wild-type (WT) mice, 5xFAD mice and non-carrier controls (MMRRC stock #34840) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA). The 5xFAD (AD) (B6SJL-Tg (APPSwFlLon, PSEN1* M146L* L286V) 6799Vas/Mmjax) mouse is a double transgenic APP/PS1 mouse model that co-expresses five AD mutations leading to accelerated plaque formation and increased Aβ42 production. This AD mouse model over-expresses APP with K670N/M671L (Swedish Mutation), I716V (Florida mutation), and V717I (London mutation), PS1 with M146L and L286V mutations. These mice accumulate high levels of intra-neuronal Aβ-42 around 1.5 months of age with amyloid deposition around 2 months (27 (link), 28 (link)). The adult Atg5-/- mice were obtained from Herbert W. Virgin from Washington University with permission of Noboru Mizushima (29 (link)). All animal experiments were performed according to protocols approved by the Animal Care and Use Committee (IACUC) of the Ohio State University College of Medicine.
7
5XFAD Mouse Model of Alzheimer's
8
Alzheimer's Disease Transgenic Mouse Model
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Transgenic mouse (strain name; B6SJL-Tg(APPSwF1Lon, PSEN1*M146L*L286V) 6799Vas/Mmjax) carrying five mutations associated with early onset familial Alzheimer’s disease (5XFAD) was used throughout this experiment. The 5XFAD mice were acquired from Jackson Laboratory (Maine, USA) and have been conserved through mating with C57BL/6 X SJL wild type mice. All mice were bred in a laboratory animal breeding room at Yonsei University (Seoul, South Korea) under regulated conditions with 12hour/12hour light-dark phase. Food and Water were provided ad libitum. All animal experiments were conducted in accordance with the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals. The research protocols were authorized by the Institutional Animal Care and Use Committee of Yonsei University.
9
Analyzing Alzheimer's Pathogenesis in 5xFAD Mice
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All experimental procedures followed protocols approved by the bioethics committee of Universidad de Santiago de Chile (report #457), following the international guidelines on animal handling and manipulation and the Chilean National Agency for Research and Development (ANID) bioethics and biosecurity standards. All experiments and methods were conducted in accordance with the ARRIVE guidelines. 5xFAD mice (Jackson laboratory, Bar Harbor, Maine, USA) and WT mice (B6SJLF1/J) were used in this study21 (link). These animals were maintained in Universidad de Valparaíso, with a time routine of 12:12 hrs light/dark cycle, with controlled temperature, and water and food ad libitum. The animals were grouped into young subjects (2–3 months-old), age at which the animals start exhibiting amyloid- brain aggregation, and adult subjects (6–7 months-old), which present thorough accumulation of plaques in the brain and behavior alterations42 (link).
10
5XFAD Mouse Model of Alzheimer's
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5XFAD mice were purchased from Jackson Laboratory (stock number: 034840-JAX, Bar Harbor, ME, USA). Mice were housed in plastic containers under constant temperature (23 ± 1 °C) and humidity (60 ± 10%), in a 12-h light/dark cycle with free access to food and water. Five-month-old 5XFAD female mice (22 ± 2 g) and male mice (30 ± 2 g) were injected intraperitoneally with 30 mg/kg of TCA in saline (0.9% NaCl) per day for eight weeks. It was also applied to WT female and male mice of the same age. In each group, one third was female and the rest were male. All animal studies were performed in accordance with the “Principles of Laboratory Animal Care” (National Institutes of Health publication number 80–23, revised 1996) and approved in 2018 by the Animal Care and Use Guidelines Committee of Kyung Hee University (approval number: KHUASP(SE)-17-126-1).
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